Sandeep R. Pai

Free amino acid profiling in grain amaranth using LC-MS/MS.

Reliable and reproducible techniques for identification and quantification of amino acids usually require derivatization. However, techniques such as LC-MS/MS may perhaps sideline the derivatization with significant accuracy. Understanding the interaction between free amino acids and several physiological processes apparently requires that the pool be studied in the seed itself. Data on free amino acid content of grain amaranth is very inadequate. The present analysis revealed a higher concentration of essential amino acids especially Threonine and Tryptophan than non essential amino acids. The study constitutes first report of use of LC-MS/MS method for analysing the free amino acid composition of amaranth grain.

Chemical Characterization, Mineral Analysis, and Antioxidant Potential of Two Underutilized Berries (Carissa carandus and Eleagnus conferta) from the Western Ghats of India

Diversity needs to be “conserved through use.” Owing to the global need, the present study includes two underutilized berries, Carissa carandus and Eleagnus conferta, to explore their potential as supplementary food by evaluating their antioxidant activity and approximate chemical composition. Berries of E. conferta are a richer source of carotene, ascorbic acid, protein, and magnesium, whereas berries of C. carandus are rich in sugars, lipids, iron, and magnesium. The antioxidant properties of the berries and a comparative account on their performance using DPPH and FRAP assay was studied. Both assays revealed high activity in the peel extract of berries of C. carandus. The LC-MS/MS profile of C. carandus reveals the presence of eleven different polyphenolic compounds while E. conferta had only four of these compounds.

Seasonal variation in content of camptothecin from the bark of Nothapodytes nimmoniana (Grah.) Mabb., using HPLC analysis

Objective: To study and compare seasonal variation in camptothecin (CPT) content from bark samples of Nothapodytes nimmoniana obtained from geographically and climatologically isolated populations. Methods: A standard High Performance Liquid Chromatography methodology was used to analyze and quantify CPT from bark samples of N. nimmoniana. Results: Sample collected from Amboli yielded highest CPT content 1.337 g/100 g dry bark powder during the monsoon compared to other localities in study. Monsoon (August) showed to accumulate higher levels of CPT in barks of N. nimmoniana as compared to summer (May). Amboli averaged highest accumulation of CPT compared to other localities under study. Conclusion: These findings indicate season to have control over accumulation of CPT. Locality Amboli has highest CPT content in all seasons and were the elite population during the study. The study also suggests the need for further investigation in lights of biosynthesis in the plant.

Phenolic contents and antioxidant properties from aerial parts of achyranthes coynei sant

Aim of the study was to evaluate antioxidant activity and total phenolic content of Achyranthes coynei; an endemic plant used in treatment of several diseases in the same lines that of Achyranthes aspera by traditional practitioners of Belgaum region. Efficiency of extraction methods was studied for aerial parts (leaves, stem, and inflorescence) extracted in methanol using continuous shaking, microwave assisted and ultra sonic extraction technique, by exposing it for different time period. Total phenolic content was measured by Folin-Ciocalteu method and antioxidant activity using 2,2’-diphenyl-1-picryl hydrazyl radical scavenging assay and ferric reducing antioxidant power assay. Extracts of A. coynei revealed highest yield of total phenolic content in continuous shaking method compared to other methods. Significantly higher amount of phenolic content (467.07±23.35 tannic acid equivalent and 360.83±18.04 caffic acid equivalent mg/100 g FW) was estimated at 360 min of continuous shaking extraction. In 2,2’-diphenyl-1-picryl hydrazyl radical scavenging assay and ferric reducing antioxidant power assay, inflorescence and leaf showed highest potential activity, respectively. Stem extracts showed lower yield of total phenolic content and antioxidant activity. Results also showed 2,2’-diphenyl-1-picryl hydrazyl radical scavenging assay had significant correlation with total phenolic content. This is first report of total phenolic content and antioxidant studies in A. coynei.

RP-HPLC analysis of seco-iridoid glycoside swertiamarin from different Swertia species

Abstract Genus Swertia is valued for its great medicinal potential; mainly Swertia chirayita (Roxb. ex Fleming) H. Karst. is used in traditional medicine for a wide range of diseases. Seco-iridoid glycosides like swertiamarin is referred with enormous pharmacological potentials. The aim of the study was to identify a suitable substitute to S. chirayita by quantifying seco-iridoid swertiamarin from five different Swertia species endemic to the Western Ghats. The reverse-phase high-performance liquid chromatography diode array detector analyses were performed and chromatographic separation was achieved on a Lichrospher 100, C18e (5 µm) column (250–4.6 mm). A mobile phase consisting of acetonitrile and water (25:75) was used for separation. Results indicated that the concentration of the marker compound has been found to vary largely between and within the species from different localities. The content of swertiamarin was the highest in S. chirayita compared to the other species studied herein, advocating the use of Swertia minor as an alternate source to S. chirayita. Graphical abstract

New Report of Triterpenoid Betulinic Acid along with Oleanolic Acid from Achyranthes aspera by Reversed-Phase-Ultra Flow Liquid Chromatographic Analysis and Confirmation Using High-Performance Thin-Layer Chromatographic and Fourier Transform-Infrared Spectroscopic Techniques

Triterpenoid betulinic acid (BA) was detected, quantified, and reported for the first time from leaf extract of Achyranthes aspera along with much known oleanolic acid (OA). Extraction was achieved using ultrasonic exposure, and reversed-phase.ultra flow liquid chromatographic (RP.UFLC) technique was employed during investigation. RP.UFLC separation was achieved on a Hibar 250–4.6 mm, 5 μ, Lichrospher 100, C18e column using methanol and water (90:10) as mobile phase with pH adjusted to 5.0 using glacial acetic acid (GAA) in an isocratic mode. The content of BA (0.25 mg g−1 fresh weight [FW]) was ∼75% higher than OA (0.06 mg g−1 FW). These results suggest BA to be the major triterpenoid compared to OA in the leaf of A. aspera. High-performance thin-layer chromatography (HPTLC) separation of the two triterpenic acids (oleanolic and betulinic acid) was also achieved on silica gel G 60 F254, 50 × ~ 100 mm glass TLC plates, using benzene, ethyl acetate, and formic acid as solvent system in a ratio of 67.9:22.7:9.4. Anisaldehyde reagent was used for detection. The method was used for the screening of oleanolic and betulinic acids from A. aspera leaf extract. Similarly, Fourier transform-infrared (FT-IR) spectroscopic analysis was done in the mid IR region of 400–4000 cm−1 with 64 scan speed using OMNIC 8.1 (ver. 8.1.210) software. The results were also supported by HPTLC and FT-IR data.

In vitro antimicrobial activity of Achyranthes coynei Sant.

Objective To validate the traditional use of Achyranthes coynei (A. coynei) Sant. as an antimicrobial in treatment of various infectious diseases.

Compound Specific Extraction of Camptothecin from Nothapodytes nimmoniana and Piperine from Piper nigrum Using Accelerated Solvent Extractor

Effects of varying temperatures with constant pressure of solvent on extraction efficiency of two chemically different alkaloids were studied. Camptothecin (CPT) from stem of Nothapodytes nimmoniana (Grah.) Mabb. and piperine from the fruits of Piper nigrum L. were extracted using Accelerated Solvent Extractor (ASE). Three cycles of extraction for a particular sample cell at a given temperature assured complete extraction. CPT and piperine were determined and quantified by using a simple and efficient UFLC-PDA (245 and 343 nm) method. Temperature increased efficiency of extraction to yield higher amount of CPT, whereas temperature had diminutive effect on yield of piperine. Maximum yield for CPT was achieved at 80°C and for piperine at 40°C. Thus, the study determines compound specific extraction of CPT from N. nimmoniana and piperine from P. nigrum using ASE method. The present study indicates the use of this method for simple, fast, and accurate extraction of the compound of interest.

Quantitative determination of three pentacyclic triterpenes from five Swertia L. species endemic to Western Ghats, India, using RP-HPLC analysis

Aim of this study was to identify pentacyclic triterpenoids betulinic acid (BA), oleanolic acid (OA) and ursolic acid (UA) from five Swertia species endemic to Western Ghats, which are used as substitutes as well as adulterants to Swertia chirayita. Our results indicate that the concentration of active compound vary largely among and within the species from different localities. OA was found in a higher amount in all species compared to BA and UA. From the results, it was clear that BA, OA and UA are present in the endemic species collected from Western Ghats, thus advocating the use of these species as alternate sources to S. chirayita. This in due course may release pressure of exploitation from natural resources of S. chirayita and help to bring it out from an endangered category from conservation point of view.

Monitoring seasonal variation of epicatechin and gallic acid in the bark of Saraca asoca using reverse phase high performance liquid chromatography (RP-HPLC) method

Background: Saraca asoca (Roxb.) Wilde (Fabaceae) is a high valued but vulnerable medicinal plant of Western Ghats region. This plant is mainly known for its use in various gynecological disorders. Objective: The objective of the present study was to investigate seasonal variation of the polyphenolic compounds viz., epicatechin and gallic acid in the bark of S. asoca by using Reverse Phase High Performance Liquid Chromatography-Diode Array Detector (RP-HPLC-DAD) method. Materials and Methods: The bark was collected in six different Ritu (season) viz. Varsha (monsoon), Sharad (autumn), hemant (early winter), Shishir (winter), Vasanta (spring), and Grishma (summer) mentioned in Ayurveda. Results : The RP-HPLC-DAD analysis indicated that levels of epicatechin and gallic acid in the bark of S. asoca vary seasonally. The highest concentration of epicatechin was observed in Shishir Ritu (3315.19 ± 165.76 mg/100g) and gallic acid during Hemant Ritu (211.90 ± 10.60 mg/100 g). Conclusions: In present study, the ability to synthesize and accumulate both the compounds in bark of S. asoca varied greatly throughout the seasons. It was also observed that the compound epicatechin was present abundantly as compared to gallic acid throughout the seasons.